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醋龟甲配方颗粒的位点特异性PCR鉴别

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目的:建立一种醋龟甲配方颗粒特异性位点聚合酶链式反应(PCR)鉴别方法.方法:根据乌龟Chinemys reevesii(Gray)及其混伪品的细胞色素C氧化酶亚基Ⅰ基因序列上的单核苷酸多态性位点设计醋龟甲的特异性鉴别引物,经过优化反应体系,确定最佳的PCR反应条件,并对该方法进行耐受性和适用性考察.结果:当退火温度为 62℃、循环数为 34 时,醋龟甲饮片、标准汤剂冻干粉和配方颗粒均可在约 92 bp处得到清晰单一的特异性条带,其混伪品及阴性对照均无条带.结论:建立的位点特异性PCR鉴别方法可快速、准确地鉴别醋龟甲饮片、标准汤剂冻干粉和配方颗粒的真伪,为醋龟甲配方颗粒质量标准研究提供了参考.
Identification of Vinegar Testudinis Carapax et Plastrum Formula Granules by Allele-Specific PCR
Objective:To establish an allele-specific polymerase chain reaction(PCR)identification method for vinegar Testudinis Carapax et Plastrum formula granules.Methods:Based on the single nucleotide polymorphism on the cytochrome C oxidase subunit Ⅰgene sequence of Chinemys reevesii(Gray)and its adulterant,specific primers of vinegar Testudinis Carapax et Plastrum were designed.The optimal PCR reaction system was determined by optimizing the reaction system,and the tolerability and applicability of this method were investigated.Results:When the annealing temperature was 62℃,and the cycle number was 34,the specific band of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules could be obtained at about 92 bp,while no band was found in other adulterants and negative control.Conclusion:The established allele-specific PCR identification method can quickly and accurately identify the authenticity of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules,providing a reference for the research on quality standards of vinegar Testudinis Carapax et Plastrum.

vinegar Testudinis Carapax et Plastrumformula granulesallele-specific PCRmolecular identification

谭斯尹、潘礼业、宋叶、刘闪闪、范耀耀、罗宇琴、李国卫

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广东一方制药有限公司,广东 佛山 528244

广东省中药配方颗粒企业重点实验室,广东 佛山 528244

醋龟甲 配方颗粒 位点特异性聚合酶链式反应 分子鉴定

工业和信息化部2022年产业技术基础公共服务平台项目2022年佛山市南海区重点领域科技攻关专项

2022-230-221南科[2023]20号-18

2024

中国现代中药
中国中药协会,中国医药集团总公司,中国药材公司

中国现代中药

CSTPCD
影响因子:0.65
ISSN:1673-4890
年,卷(期):2024.26(8)